Heparin Binds Endothelial Cell Growth Factor, the Principal Endothelial Cell Mitogen in Bovine Brain

Abstract

Endothelial cell growth factor (ECGF), an anionic polypeptide mitogen, binds to immobilized heparin. The interaction between the acidic polypeptide and the anionic carbohydrate suggests a mechanism that is independent of ion exchange. Monoclonal antibodies to purified bovine ECGF inhibited the biological activity of ECGF in crude preparations of bovine brain. These data indicate that ECGF is the principal mitogen for endothelial cells from bovine brain, that heparin affinity chromatography may be used to purify and concentrate ECGF, and that the affinity of ECGF for heparin may have structural and perhaps biological significance.

Purification of a factor from human placenta that stimulates capillary endothelial cell protease production, DNA synthesis, and migration.

“…It is not clear how this molecule is related to other endothelial cell mitogens that have been isolated. It has been proposed that the known endothelial cell mitgens fall into two classes: molecules related to endothelial cell growth factor, which are eluted from heparin-Sepharose with 1 M NaCl and have an acidic pI (8)(9)(10), and molecules related to fibroblast growth factor, which bind more strongly to heparin-Sepharose and have a basic pI (7,10,12). Molecules from each ofthese classes have been shown to be angiogenic in vivo (22,23).…”

“…3B shows that addition of the protease-inducing factor to cultures of BCE cells stimulated the incorporation of "25I-labeled deoxyuridine into DNA in a dose-dependent manner. Stimulation of 125"-labeled deoxyuridine incorporation was achieved with the same concentrations of factor that were able to induce production of PA have been purified as endothelial cell mitogens (7)(8)(9)(10)(11)(12). We have previously determined that, with crude placenta sonicate, increased incorpration of '25I-labeled deoxyuridine into DNA correlates with other measurements of mitogenesis (21).…”

“…However, it has not been demonstrated whether the stimulation of these different processes is due to a single molecule in the preparations or to several different molecules. Several groups have used heparin-Sepharose affinity chromatography to purify endothelial cell mitogens from various sources (7)(8)(9)(10)(11)(12). We have investigated whether heparin-Sepharose affinity chromatography could also be used to purify the PA and collagenase-inducing activity from a preparation with known angiogenic activity and whether the purified proteaseinducing molecule was also able to induce the other activities associated with angiogenesis-increased endothelial cell mitosis and motility.…”

Fibroblast growth factor/fibroblast growth factor receptor system in angiogenesis

“…TAF had a molecular weight of about 10 kDa and consisted of 25% RNA, 10% proteins, and 58% carbohydrates, plus a possible lipid fraction. The 1980s saw for the first time the purification to homogeneity of pro-angiogenic proteins, the breakthrough coming as a result of the observation that endothelial cell growth factors showed a marked affinity for heparin [6,7]. This led to the identification, purification, and sequencing of the two prototypic heparin-binding angiogenic growth factors FGF1 and FGF2.…”

Chemical and biological characterization of a truncated form of acidic fibroblast growth factor from bovine brain

“…Furthermore, a variety of biochemical and biological data strongly suggest that less well characterized growth factors, e. g. from placenta [15], macrophages [16], a transplantable chondrosarcoma 1171, brain [18 -201, eye or retina [18,21,22] and cartilage [23] are structurally and biologically similar if not identical to either bFGF or aFGF. Likewise, growth factors with other names, e. g. endothelial cell growth factor (ECGF) [24] or heparin-binding growth factors [25] are now thought to be identical to one or the other of the FGF forms [19, 261. bFGF is found in at least two molecular forms: upon purification, bovine pituitary and brain yield a 146-residue protein as characterized [l, 2, 101. In contrast, the bFGF form found in kidney or corpus luteum tissue is amino-terminally truncated (the 15 amino-terminal residues are.…”